Project Summary Asthma is a chronic lung disease that causes the airways in the lungs to become inflamed, making it difficult to breathe and leading to episodes of intense coughing and wheezing. The prevalence of asthma has increased rapidly over the past several decades, and more than 1 in 12 Americans are now living with the disease. While there is no cure for asthma, the symptoms of the disease can be managed through a series of prescription medicines. However, this conventional, one-size-fits-all therapeutic approach fails to account for the different clinical forms and phenotypes of asthma, which have been the subject of many recent medical studies. By analyzing the different cell populations found in sputum, the mucus within the airways of the lungs, researchers have identified the distinct immunological phenotypes associated with the disease. Identifying these phenotypes has led to hopes of developing individually tailored therapeutic treatments that will more effectively target the mechanisms unique to each phenotype. Although sputum analysis has proven to be a powerful tool that provides a noninvasive means of characterizing the different variations of asthma, the current methods for processing and analyzing sputum are complex and labor-intensive. The multi-step process requires highly trained personnel, and the amount of sputum collected from a patient is often too small to perform meaningful analysis. In addition, the process requires the use of expensive, benchtop equipment, which prevents point-of- care applications and limits the analysis to centralized facilities. As a result, there exists a critical need in the medical community for a more simple and rapid approach for processing and analyzing low-volume sputum samples. In this SBIR project, we will address this unmet need by developing and commercializing acoustofluidic (i.e., the fusion of acoustics and microfluidics) technologies for point-of-care, automated sputum processing and analysis. In Phase I, Ascent has successfully demonstrated the utility and feasibility of the proposed devices by meeting or exceeding the acceptable values of each of the five key parameters identified in the Measures of Success. In Phase II, our commercialization activities will improve performance of the disposable acoustofluidic chips, develop self-contained, beta-testing-ready prototypes, and validate their performance with a pilot clinical feasibility study. The proposed system will have the following features: 1) ability to perform accurate sputum analysis over a much wider sample size range (volume: 50?3,000 L) than the conventional approaches (volume: 1,000?3,000 L); 2) automation and low turnaround time; 3) biohazard containment; and 4) low-cost, point-of-care devices. With these features, we expect that once demonstrated, the proposed acoustofluidic platform will not only be an excellent replacement for existing sputum processing/analysis tools, but will also fulfill many unmet needs for applications where the amount of sputum induced from asthmatic patients is not enough to run the standard tests and/or the expertise and equipment to perform this analysis are not available, such as most practice locations outside of large hospitals.